Adenylyl cyclase signaling in persistent pain

持续性疼痛中的腺苷酸环化酶信号传导

基本信息

批准号：
10418657
负责人：
DEREK C MOLLIVER
金额：
$ 45.99万
依托单位：
UNIVERSITY OF NEW ENGLAND
依托单位国家：
美国
项目类别：
财政年份：
2019
资助国家：
美国
起止时间：
2019-08-01 至 2024-04-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10418657
关键词：
A kinase anchoring protein Acute Acute Pain Adenylate Cyclase Affect Afferent Neurons American Analgesics Animals Arthritis Automobile Driving Behavior assessment Behavioral Biochemical Brain CRISPR/Cas technology Cells Cessation of life Chronic Clinical Co-Immunoprecipitations Complex Coupled Cyclic AMP Cyclic AMP-Dependent Protein Kinases Dangerousness Development Diabetic Neuropathies Dinoprostone Electrophysiology (science)Family Family member G Protein-Coupled Receptor Signaling Gene Deletion Genes Goals Hyperalgesia Hypersensitivity Image Immunohistochemistry In Vitro Inflammatory Injury Intervention Intractable Pain Investigation Ion Channel Ion Channel Gating Knock-out Knockout Mice Lead Malignant Neoplasms Mass Spectrum Analysis Mechanics Mediating Modeling Molecular Mus Neurogenic Inflammation Neurons Neuropathy Neuropeptides Nociception Nociceptors Opioid Opioid Analgesics Opioid Receptor Pain Pain management Pathologic Peripheral Persistent pain Pharmaceutical Preparations Pharmacology Phosphoproteins Phosphorylation Play Postherpetic neuralgia Pre-Clinical Model Production Protein Isoforms Receptors, Adrenergic, alpha-2 Regulation Reporter Resistance Role Scaffolding Protein Second Messenger Systems Signal Pathway Signal Transduction Signaling Protein Spinal Cord Spinal Ganglia Sustainable Development Syndrome System TRPV1 gene Testing Western Blotting allodynia base chronic pain chronic pain management clinically significant effective intervention effective therapy experimental study improved in vivo innovation non-opioid analgesic novel pain chronification patch clamp prescription opioid receptor recruit response scaffold societal costs transmission process

项目摘要

Chronic pain affects more than 50 million Americans per year, resulting in extraordinary personal and societal costs. Adding to the dilemma, deaths involving prescription opiate analgesics have almost quadrupled in the last ten years. The clinical challenge of pain management is underscored by evidence that chronic pain is mechanistically distinct from acute pain, therefore a thorough understanding of the molecular and cellular mechanisms underlying the transition to chronic pain is fundamental to improving and expanding treatment options. Hyperalgesic priming is a compelling model of the transition to chronic pain in which an initial injury resolves, but leaves the animal in a primed state in which a second insult induces a greatly prolonged pain response. While previous studies have explored the development of sustained mechanical hypersensitivity, we have adapted this model to examine sustained hypersensitivity mediated by nociceptors expressing the heat-gated ion channel TRPV1, which can drive pain in a range of inflammatory and neuropathic conditions. Experiments proposed here will test the contribution to heat hyperalgesic priming of adenylyl cyclase isoform AC2, which has not been previously characterized in sensory neurons, and which is insensitive to inhibition by Gi/o-coupled receptors such as opioid receptors. Preliminary analysis indicates that AC2 is highly expressed in TRPV1-expressing neurons and required for the manifestation of heat hyperalgesic priming. Specific Aim 1 will examine the impact of AC2 gene deletion and pharmacological inhibition on behavioral nociceptive thresholds at baseline, in acute hyperalgesia and in the setting of hyperalgesic priming. Specific Aim 2 will determine whether AC2 and the downstream effector Epac are functionally coupled through AKAP family member scaffolding proteins by co-immunoprecipitation-mass spectrometry, and will determine the impact of deleting identified AKAP genes on Epac signaling and nociceptor function in vitro using a CRISPR/Cas9-based approach. Specific Aim 3 will determine the consequences for Epac function of AC2 and AKAP gene deletion through Epac-dependent PKC phospho- substrate profiling and behavioral assessment of heat hyperalgesia. This proposal will use innovative approaches to explore novel mechanisms contributing to the development of persistent hyperalgesia.

慢性疼痛每年影响超过 5000 万美国人，导致非同寻常的后果个人和社会成本。更糟糕的是，涉及处方阿片类药物的死亡过去十年中镇痛药几乎增加了四倍。疼痛的临床挑战有证据表明，慢性疼痛在机制上与急性疼痛不同，这强调了治疗的重要性疼痛，因此要彻底了解潜在的分子和细胞机制向慢性疼痛的过渡对于改善和扩大治疗选择至关重要。痛觉过敏启动是向慢性疼痛转变的一个令人信服的模型，其中初始伤害会消退，但会让动物处于准备状态，在这种状态下，第二次伤害会引发大大延长了疼痛反应。虽然之前的研究已经探索了持续的机械超敏反应，我们已经采用这个模型来检查持续的机械超敏反应由表达热门控离子通道 TRPV1 的伤害感受器介导的超敏反应，可以引起一系列炎症和神经病性疾病的疼痛。提议的实验这里将测试腺苷酸环化酶亚型 AC2 对热痛觉过敏引发的贡献，以前没有在感觉神经元中表征过，并且对 Gi/o偶联受体（例如阿片受体）的抑制。初步分析表明 AC2 在表达 TRPV1 的神经元中高度表达，并且是表现的必需的热痛觉过敏启动。具体目标 1 将检查 AC2 基因缺失的影响和对基线行为伤害性阈值的药物抑制，急性痛觉过敏和痛觉过敏启动的情况下。具体目标 2 将决定是否 AC2 和下游效应器 Epac 通过 AKAP 家族成员进行功能耦合通过免疫共沉淀-质谱法构建支架蛋白，并确定删除已识别的 AKAP 基因对体外 Epac 信号传导和伤害感受器功能的影响使用基于 CRISPR/Cas9 的方法。具体目标 3 将决定后果通过 Epac 依赖性 PKC 磷酸化实现 AC2 和 AKAP 基因删除的 Epac 功能热痛觉过敏的基质分析和行为评估。本提案将使用探索有助于发展的新机制的创新方法持续性痛觉过敏。